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Integration position-independent expression of human globin transgenes in transgenic mice requires the presence of regulatory elements from the beta-globin locus control region (LCR) in the transgene construct. However, several recent studies have suggested that, while clearly necessary, such elements are not by themselves sufficient to realize this effect. In the case of the human fetal gamma-globin genes, previous results have indicated that additional regulatory information required for sheltering of gamma-globin transgene expression from position effects may reside downstream from the A gamma gene. To investigate this possibility, we established 17 lines of transgenic mice carrying constructs comprising a micro-LCR (microLCR) element, an A gamma-globin gene fragment, and a variable length of 3' sequence information beyond the A gamma 3' HindIII site. gamma-Globin expression during development was studied in 170 individual F2 progeny from these lines. We find that gamma-globin expression becomes sheltered from position effects when the normally position-sensitive microLCR-A gamma construct is extended by 600 bp beyond the 3' HindIII site to include a previously identified regulatory sequence (the A gamma-globin enhancer), the functional significance of which in vivo had heretofore been unclear. The results suggest that the mechanism whereby an upstream LCR achieves sheltering of globin gene expression from position effects involves cooperation with a gene-proximal regulatory element distinct from the promoter region.

molecular and cellular biology

Sheltering of γ-Globin Expression from Position Effects Requires both an Upstream Locus Control Region and a Regulatory Element 3′ to the <sup>A</sup>γ-Globin Gene

Sheltering of γ-Globin Expression from Position Effects Requires both an Upstream Locus Control Region and a Regulatory Element 3′ to the Aγ-Globin Gene

Sheltering of γ-Globin Expression from Position Effects Requires both an Upstream Locus Control Region and a Regulatory Element 3′ to the ^Aγ-Globin Gene